A simple method for developing microsatellite markers using amplified fragments of inter-simple sequence repeat (ISSR)

We have developed a simple method for isolating microsatellites. Fragments flanked by one or two microsatellites were amplified using a microsatellite, primer, (AC)(10). A primer IP1 designed from the sequenced region at one end of the microsatellite and for nested PCR another primer IP2 based on the sequence between IPI and the microsatellite were prepared. These two primers were used to determine the other sequence flanking the microsatellite by a "walking" method. With this approach, we developed several microsatellite markers from Salix reinii, Pinus densiflora and Robinia pseudoacacia, respectively. The absence of enrichment processes and screening procedures makes it easier to develop microsatellite markers, and this approach provides an alternative for the development of microsatellite markers in any organism.